An effective S-adenosylmethionine and glutathione enriching yeast mutant of Candida utilis CCTCC M 209298 was first screened from plates containing 0.5 g/L of DL-ethionine by complex mutagenesis with UV and γ-ray in this study. Medium components optimization for enhanced co-production of S-adenosylmethionine and glutathione by C. utilis CCTCC M 209298 was further carried out using response surface methodology. The significant factors influencing S-adenosylmethionine and glutathione co-production were selected by Plackett-Burman design as sucrose, KH2PO4 and L-methionine, and Box-Behnken design was applied for further optimization studies. Based on these approaches, the optimized concentrations on medium components for higher co-production of S-adenosylmethionine and glutathione were sucrose 35.4 g/L, (NH4)2SO4 10 g/L, KH2PO4 12.3 g/L, MgSO4·7H2O, 0.05 g/L, CaCl2 0.05 g/L and L-methionine 4.6 g/L. The medium optimization by response surface methodology led to a total production of 589.3 mg/L on S-adenosylmethionine and glutathione, which was 2.4-fold increased compared with the medium without optimization.